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Modification method for cell culture product in adhered state

A cell culture, adhesion state technology, applied in biochemical equipment and methods, biological preparations for removing bad cells, animal cells, etc., to achieve the effect of eliminating time, reducing burden, and improving quality

Active Publication Date: 2019-01-04
TERUMO KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, there has been no report so far on the technique of further modifying the cell culture formed into a sheet after forming a sheet.

Method used

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  • Modification method for cell culture product in adhered state
  • Modification method for cell culture product in adhered state

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0127] Example 1. Alteration test of sheet cell culture

[0128] (1) Preparation of sheet cell culture

[0129] Using skeletal muscle myoblasts (including fibroblasts) prepared from human skeletal muscle by a conventional method, a sheet-shaped cell culture was prepared. to become 3.7×10 6 In the manner of one per well, the cell mixture of human skeletal muscle myoblasts and human fibroblasts suspended in DMEM / F12 medium (Thermo Fisher Scientific Inc.) containing 20% ​​human serum was seeded on a temperature-responsive culture dish (UpCell(R) 12-well multiwell culture plate, CellSeed), at 37°C, 5% CO 2 12 to 26 hours for sheet culture. After the sheet culture, the culture medium was removed, 700 μL of cooled HBSS(+) (Thermo Fisher Scientific Inc.) was added, and allowed to stand for 10 minutes, and then the sheet cell culture was completely detached by blowing gently.

[0130] After the sheet-shaped cell culture was completely peeled off, the HBSS(+) was removed, and new...

example 2

[0134] Example 2. Alteration assay in cell suspension

[0135] Add 1.55 mL of HBSS(+) to the mixture of human skeletal muscle myoblasts and human fibroblasts in an unflapped state, and let it stand under refrigerated conditions at 2-8°C. The purity of skeletal muscle myoblasts was measured in the same manner as in Example 1 (2) above at the start of the test and 72 hours later.

[0136] show the result in figure 2 . Changes in the purity of skeletal muscle myoblasts could not be confirmed in the case of non-sheeting.

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Abstract

The present invention addresses the problem of providing a method whereby, after the formation of a cell culture product in an adhered state, the cell culture product in an adhered state is modified so as to change the content ratio of cells constituting the same. To solve this problem, provided is a method which comprises immersing a cell culture product in an adhered state, said cell culture product containing at least two kinds of cells, in an oligotrophic isotonic solution.

Description

technical field [0001] The present invention relates to a method of modifying a cell culture in an adherent state, a method of producing a sheet-like cell culture including a step of implementing the modification, and the like. Background technique [0002] In recent years, attempts have been made to transplant various cells in order to repair damaged tissues and the like. For example, in order to repair myocardial tissue damaged by ischemic heart disease such as angina pectoris and myocardial infarction, dilated cardiomyopathy, etc., attempts are being made to use fetal cardiomyocytes, skeletal muscle myoblasts, mesenchymal stem cells, cardiac stem cells, ES Cells and the like (Non-Patent Documents 1 to 2). [0003] As part of the above-mentioned attempts, a sheet-like cell culture in which cells are formed into a sheet using a cell structure formed of a scaffold has been developed (Patent Document 1, Non-Patent Document 2). [0004] Regarding the application of sheet cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N5/077
CPCC12N1/00C12N5/0081C12N5/0658C12N5/0656A61L27/3804C12N5/0087
Inventor 大山贤二竹内凉平竹内稔和
Owner TERUMO KK